Effects of decontamination, DNA extraction and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli ulcer)

We compared two DNA extraction methods (a semi-automated method using Maxwell(R) kit and a modified Boom method) and three amplification procedures (a single-step PCR, a nested PCR and a real-time quantitative PCR) on 74 surgical tissue specimens from clinically suspected Buruli ulcer patients. All these procedures were compared before and after decontamination.We observed that, among the procedures tested, real-time PCR after the modified Boom extraction method or a single-run PCR assay after the Maxwell(R) 16 extraction method, performed on non-decontaminated suspensions are the best for the molecular diagnosis of Mycobacterium ulcerans disease.

Publication year:2012

Keywords:Bacterial diseases, Buruli ulcer, Mycobacterium ulcerans, Molecular diagnostic techniques, Decontamination, DNA extraction, Amplification, Polymerase chain reaction, PCR, Comparison, Methodology, Smear-positive, Laboratory techniques and procedures